製品の概要

  • 製品名Anti-CENPF antibody [14C10 /1D8]
    CENPF 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [14C10 /1D8] to CENPF
  • アプリケーション適用あり: ICC/IF, WB, IP, IHC-P, IHC-Fr, Flow Cytmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Fusion protein, corresponding to amino acids 1759-2093 of Human Mitosin/CENPF.

  • ポジティブ・コントロール
    • HeLa
  • 特記事項

    Clone 1D8 recognizes mitosin, a 350-kDa phosphoprotein, that is specifically involved in mitotic-phase progression. Mitosin is expressed throughout S, G2, and M phases of the cell cycle but is absent in G0 and G1.

    This antibody could be of use as a proliferation marker. This protein associates with the centromere-kinetochore complex. The protein is a component of the nuclear matrix during the G2 phase of interphase. In late G2 the protein associates with the kinetochore and maintains this association through early anaphase. It localizes to the spindle midzone and the intracellular bridge in late anaphase and telophase, respectively, and is thought to be subsequently degraded. The localization of this protein suggests that it may play a role in chromosome segregation during mitotis. It is thought to form either a homodimer or heterodimer. Autoantibodies against this protein have been found in patients with cancer or graft versus host disease.

製品の特性

  • 製品の状態Liquid
  • 保存方法Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • バッファーPhosphate buffered saline
  • Concentration information loading...
  • 精製度Immunogen affinity purified
  • 特記事項(精製)1 ml of undiluted tissue culture supernatant with 15 mM sodium azide; or, 1 mg/ml of antibody purified from ascities fluid by Protein G chromatography to at least 95% homogeneity as determined by SDS-PAGE. Prepared in 10mM PBS (pH 7.4).
  • ポリ/モノモノクローナル
  • クローン名14C10 /1D8
  • ミエローマNS1
  • アイソタイプIgG2a
  • 軽鎖の種類kappa
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab90 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 367 kDa.
IP Use at an assay dependent concentration.
IHC-P 1/200.
IHC-Fr Use at an assay dependent concentration.
Flow Cyt Use 0.5µg for 106 cells.

ターゲット情報

  • 機能Required for kinetochore function and chromosome segregation in mitosis. Required for kinetochore localization of dynein, LIS1, NDE1 and NDEL1. Regulates recycling of the plasma membrane by acting as a link between recycling vesicles and the microtubule network though its association with STX4 and SNAP25. Acts as a potential inhibitor of pocket protein-mediated cellular processes during development by regulating the activity of RB proteins during cell division and proliferation. May play a regulatory or permissive role in the normal embryonic cardiomyocyte cell cycle and in promoting continued mitosis in transformed, abnormally dividing neonatal cardiomyocytes. Interaction with RB directs embryonic stem cells toward a cardiac lineage. Involved in the regulation of DNA synthesis and hence cell cycle progression, via its C-terminus. Has a potential role regulating skeletal myogenesis and in cell differentiation in embryogenesis. Involved in dendritic cell regulation of T-cell immunity against chlamydia.
  • 関連疾患Stromme syndrome
  • 配列類似性Belongs to the centromere protein F family.
  • 発生段階Gradually accumulates during the cell cycle, reaching peak levels in G2 and M phase, and is rapidly degraded upon completion of mitosis.
  • 翻訳後修飾Hyperphosphorylated during mitosis.
  • 細胞内局在Cytoplasm, perinuclear region. Nucleus matrix. Chromosome, centromere, kinetochore. Cytoplasm, cytoskeleton, spindle. Relocalizes to the kinetochore/centromere (coronal surface of the outer plate) and the spindle during mitosis. Observed in nucleus during interphase but not in the nucleolus. At metaphase becomes localized to areas including kinetochore and mitotic apparatus as well as cytoplasm. By telophase, is concentrated within the intracellular bridge at either side of the mid-body.
  • Information by UniProt
  • 参照データベース
  • 別名
    • AH antigen antibody
    • Cell cycle dependent 350K nuclear protein antibody
    • CENF antibody
    • CENP F antibody
    • CENP-F antibody
    • CENPF antibody
    • CENPF kinetochore protein antibody
    • CENPF_HUMAN antibody
    • Centromere protein F 350/400ka antibody
    • Centromere protein F antibody
    • Hcp 1 antibody
    • Hcp1 antibody
    • Kinetochore protein CENP F antibody
    • Kinetochore protein CENPF antibody
    • Mitosin antibody
    • PRO1779 antibody
    see all

Anti-CENPF antibody [14C10 /1D8] 画像

  • ab90 (1/200) staining CENPF in HeLa cells (green). Cells were fixed in paraformaldehyde, permeabilised with 0.5% Triton X100/PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • Overlay histogram showing HeLa cells stained with ab90 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab90, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • CENPF was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to CENPF (ab90)and 50µl of protein G magnetic beads (lane 1). The antibody was incubated with the Protein G beads for 10min under agitation. No antibody was added to the control (lane 2). Hela whole cell extract diluted in RIPA buffer was added to each sample and incubated for 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab90. Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Bands: 370kDa:CENPF; 171kDa: non specific - as present in control (lane 2); 70kDa: We are unsure as to the identity of this extra band.

Anti-CENPF antibody [14C10 /1D8] (ab90) 使用論文

This product has been referenced in:
  • Jodoin JN  et al. Nuclear-localized Asunder regulates cytoplasmic dynein localization via its role in the integrator complex. Mol Biol Cell 24:2954-65 (2013). WB ; Human . Read more (PubMed: 23904267) »
  • Groth P  et al. Homologous recombination repairs secondary replication induced DNA double-strand breaks after ionizing radiation. Nucleic Acids Res : (2012). Read more (PubMed: 22505579) »

See all 8 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Dr. Alexander Garvin

Verified customer

投稿 Jul 06 2016

Sorry for the delay in getting back to you.

I now have more information to share with you in regards to what size band would be expected when using Anti-CENPF antibody [14C10 /1D8] in western blotting.

As discussed over the phone, t...

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Thank you for providing the datasheet for Anti-CENPF antibody [14C10 /1D8] (ab90) which we sent to you.

As discussedover the phone,I am currently looking into why the information relating to the expected protein size has been changed and exac...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

投稿 Oct 14 2008

Thank you for your enquiry. I am sorry to hear that you have been having difficulties with this antibody. This is the first complaint that we have received relating to this antibody. We have received positive western blotting and immunoprecipitation...

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No, I'm afraid not. We can only guarantee that the antibody will work as stated on the online datasheet (that is, in the applications for which it has been tested). If you have any additional questions, please contact us again.

Thank you for your enquiry. To our knowledge, this antibody has yet to be tested in ICC. Ab90 has been tested for application in Western blot, Immunoprecipitation, Immunohistochemistry (Formalin-fixed paraffin-embedded sections), and Immunohistochemist...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"